WO2017071479A1

WO2017071479A1 - Glutamine acyl cyclase inhibitor

Info

Publication number: WO2017071479A1
Application number: PCT/CN2016/101995
Authority: WO
Inventors: 吴海强; 刘志刚; 贺震旦; 郑易之; 邹永东
Original assignee: 深圳大学
Priority date: 2015-10-26
Filing date: 2016-10-13
Publication date: 2017-05-04
Also published as: CN105384692B; CN105384692A

Abstract

Disclosed is a glutamine acyl cyclase inhibitor, and the structure thereof is as shown in formula (I). The glutamine acyl cyclase inhibitor (QC inhibitor) provided in the present invention is designed according to the target enzyme protein activity center crystal structure, shows good selectivity and specificity, and has a higher druggability; and the maternal structure thereof is richer, and same has a good water solubility, a good transmembrane performance, and a higher activity.

Description

Glutaminoyl cyclase inhibitor

Technical field

The invention relates to the field of medicinal chemistry, and in particular to a glutaminyl cyclase inhibitor.

Background technique

Glutaminylcyclase (QC, EC 2.3.2.5) is an enzyme that catalyzes the intramolecular cyclization of N-terminal glutamine residues such as polypeptides and proteins to form pyroglutamic acid (pGlu). In 1963, QC was first discovered in the tropical plant Carica papaya latex. Later studies confirmed that QC is distributed in plants, animals and microorganisms. Mammalian QC is highly conserved, has no sequence homology with papaya QC, and has significant sequence homology with bacterial aminopeptidase, so animal and plant QC have different evolutionary origins. In plants, the physiological function of QC is not very clear, and may play a role in the process of plant defense against pathogenic microorganisms. QC in animals has important biological functions such as changing the N-terminal chemical structure, regulating activity and enhancing stability of proteins. The key role of QC activity in the pathogenesis of many major diseases has been highly valued by people.

Alzheimer's disease (AD) is a common neurodegenerative disease and is the main form of Alzheimer's disease. More than 65% of AD patients exceed the total number of Alzheimer's patients. With the aging of the global population, the incidence of AD and the number of patients are rapidly increasing. Elevated, epidemiological surveys show that AD has become the third largest worldwide health problem after cardiovascular and cerebrovascular diseases and cancer. The main symptoms of AD include progressive memory and cognitive dysfunction. It has the characteristics of irreversible and high mortality. There is no specific treatment in clinical practice.

The pathogenesis of AD is complex, and many hypotheses have been proposed from different perspectives, including β-amyloid (Aβ) cascade theory, Tau protein entanglement theory, oxidative stress theory, biometal ion homeostasis theory, mitochondria Functional failure theory and so on. However, the exact pathological mechanism of AD is still unclear. Research on anti-AD drugs based on the above academic hypothesis is progressing slowly.

Clinical pathology studies have confirmed that AD pathological features are mainly Aβ precipitation outside the brain neurons and hyperphosphorylation of Tau protein tangles inside the neurons. However, in recent years, a number of in vivo and clinical studies have found that, unlike normal A1 senile plaque deposits in the brain, the main component of senile plaque deposition in AD patients is not Aβ, but a variant Aβ, especially N-terminal glutamine. The content of pGlu-Aβ circulated in the residue molecule, especially pGlu-Aβ42/pGlu-Aβ40, is more than 50%. pGlu-Aβ has stronger neurotoxicity, faster aggregation and sedimentation rate, stronger stability, and its production is directly related to the occurrence of clinical symptoms of AD. It is a marker that is more specific than Aβ and more specific to the pathogenesis of AD. .

Further research shows that pGlu-Aβ is a product of QC enzyme catalysis, and selective inhibition of QC activity can significantly inhibit the production of pGlu-Aβ and the formation of senile plaque, and significantly improve the symptoms of AD such as cognitive and memory impairment. . At the same time, the study found that the high expression of QC in the brain of AD patients is a landmark lesion in the early stage of AD. The characteristics of QC RNA can be detected in peripheral blood before the production of pGlu-Aβ and Aβ. Up. It can be seen that high expression of QC is a key factor in the pathogenesis and development of AD. QC has opened a new research direction for AD pathology and etiological anti-AD drug research. QC inhibitors are expected to become a strategic breakthrough for innovative anti-AD drug development.

In addition, QC is highly expressed in the pathogenesis of complex diseases such as rheumatoid arthritis, nonalcoholic hepatitis, cutaneous melanoma, and lupus erythematosus, which are related to the intrinsic immune dysfunction of the body. QC inhibitors are also useful in the treatment of a variety of diseases as described above.

Currently, there are relatively few studies related to QC inhibitors. A class of linear QC inhibitor molecules is disclosed in EP1713780B1, EP2091948B1, US7304086B2, US7371871B2, US7741354B2, US7892771B2, US8129160B2, US8188094B2, US8202897B2, US8227498B2, US20080214620A1, US200880286231A1, US20090269301A1. There are certain deficiencies in the structure and activity of such QC inhibitors, such as single matrix structure, poor water solubility, poor transmembrane performance, limited activity, and complicated synthesis steps. Therefore, further expand the chemical structure diversity of QC inhibitors, design new QC inhibitors with higher medicinal properties, and study QC inhibitors for AD, rheumatoid arthritis, non-alcoholic hepatitis, cutaneous melanoma, lupus erythematosus syndrome The therapeutic effects and mechanisms of diseases, etc., have extremely important scientific significance and application value for the study of pathological mechanism related to high expression of QC, research on etiological new drugs, early diagnosis and treatment of QC targeting.

Summary of the invention

In view of the above deficiencies of the prior art, the present invention aims to provide a glutaminyl cyclase inhibitor, which aims to solve the problem that the existing QC inhibitor has a single mother structure. Poor water solubility, poor transmembrane performance, limited activity, etc.

The technical solution of the present invention is as follows:

A glutaminyl cyclase inhibitor wherein the structural formula is as follows:

Wherein, the A unit is a benzene ring, a six-membered heteroaryl ring, a five-membered heteroaryl ring, a seven-membered aromatic ring, a naphthalene, an anthracene, a naphthoquinone or a polyaromatic ring system, and R ₁ is a hydrogen, a linear alkyl group, or a branched chain. One of an alkyl group, an alkoxy group, a halogen, a carboxyl group, a nitro group, a sulfonic acid group, an amine group, a phosphate group, and a substituent thereof;

The B unit is a benzene ring, a six-membered heteroaryl ring, a five-membered heteroaryl ring, a seven-membered aromatic ring, a naphthalene, an anthracene, a naphthoquinone or a polyaromatic ring system, and R ₂ is a hydrogen, a linear alkyl group, or a branched alkane. One of a group, an alkoxy group, a halogen, a carboxyl group, a nitro group, a sulfonic acid group, an amine group, a phosphate group, and a substituent thereof;

In the C unit, R ₃ on the imidazole ring is one of hydrogen, a linear alkyl group, a branched alkyl group, an alkoxy group, a halogen, an amine group, and a substituent thereof.

The glutaminyl cyclase inhibitor, wherein the B unit and the C unit are ortho, meta or para at the linking position of the A unit.

The glutaminyl cyclase inhibitor, wherein the parent core structure of the A unit and the B unit is the same or different in the same compound, and the structures of R ₁ and R ₂ are the same or different.

The glutaminyl cyclase inhibitor, wherein R ₁ is a single substitution or a multiple substitution at a different position.

The glutaminyl cyclase inhibitor, wherein R ₂ is a monosubstituted or polysubstituted at a different position.

The glutaminyl cyclase inhibitor, wherein R ₃ is a single substitution or a multiple substitution at a different position.

The glutaminyl cyclase inhibitor, wherein the substitution position of R ₃ is at the 2, 4 or 5 position of the imidazole ring.

The glutaminyl cyclase inhibitor, wherein, in the C unit, the carbon atom in the alkyl chain is n=1-4.

Advantageous Effects: A glutaminyl cyclase inhibitor (QC inhibitor) provided by the present invention is designed according to a crystal structure of a target enzyme protein active center, and exhibits good selectivity and specificity, and has Higher drug-forming properties; its parent structure is more abundant, water-soluble, transmembrane performance and activity higher.

DRAWINGS

Fig. 1 is a schematic view showing the principle of QC enzyme inhibitory activity test in the present invention.

Detailed ways

The present invention provides a glutaminyl cyclase inhibitor, and the present invention will be further described in detail below in order to clarify and clarify the objects, aspects and effects of the present invention. It is understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

The present invention relates to a glutaminyl cyclase inhibitor (QC inhibitor) having the following structural formula:

The QC inhibitor of the present invention comprises three pharmacophore structural units: A, B and C, wherein the A unit is a benzene ring, a six-membered heteroaryl ring (N, S, O), and a five-membered heteroaryl ring ( C, N, S, O), seven-membered aromatic ring, naphthalene, anthracene, naphthoquinone or (hetero) polyaromatic ring system, R ₁ is hydrogen, linear alkyl group, branched alkyl group, alkoxy group, Halogen, carboxyl, nitro, sulfonate, amine, phosphate, and substituents thereof (eg, hydrogen, linear alkyl, branched alkyl, alkoxy, halogen, carboxyl, nitro, sulfonate) One of a substituent of an amine group or a phosphate group; R ₁ is a single substitution or a multiple substitution at a different position.

The B unit is a benzene ring, a six-membered heteroaryl ring (N, S, O), a five-membered heteroaryl ring (C, N, S, O), a seven-membered aromatic ring, naphthalene, anthracene, naphthoquinone or (hetero) An aromatic ring system, R ₂ is hydrogen, a linear alkyl group, a branched alkyl group, an alkoxy group, a halogen, a carboxyl group, a nitro group, a sulfonic acid group, an amine group, a phosphoric acid group, and a substituent thereof (for example, hydrogen, straight One of a chain alkyl group, a branched alkyl group, an alkoxy group, a halogen group, a carboxyl group, a nitro group, a sulfonic acid group, an amine group or a phosphate group; R ₂ is a single substituent or a plurality of different positions Replace.

In the C unit, R ₃ on the imidazole ring is one of hydrogen, a linear alkyl group, a branched alkyl group, an alkoxy group, a halogen, an amine group, and a substituent thereof. R ₃ is a single substitution or a multiple substitution at a different position. The substitution position of R ₃ is at the 2, 4 or 5 position of the imidazole ring. In the C unit, the carbon atom in the alkyl chain has n = 1-4, for example, n is 1, 2, 3 or 4.

Further, the linking positions of the B unit and the C unit at the A unit are ortho, meta or para.

Further, in the same compound, the parent core structures of the A unit and the B unit are the same or different, and the structures of R ₁ and R ₂ are the same or different.

The QC inhibitors of the present invention are pharmaceutically acceptable salts, including lithium salts, sodium salts, potassium salts, magnesium salts, calcium salts, iron salts, copper salts, organic ammonium salts, hydrochloride salts, phosphate salts, acetic acid. Salt, propionate, oxalate, citrate, and the like. The QC inhibitor of the invention is a new type of QC inhibitor disclosed for the first time, and has extremely important scientific significance and research value for the research of self-directed QC targeted innovation lead drug in China. It can be widely used in high-efficiency QC inhibitors, such as QC targeted anti-AD lead drugs, treatment of QC-specific high expression-related diseases (including rheumatoid arthritis, non-alcoholic hepatitis, cutaneous melanoma, lupus erythematosus syndrome) Etc.), or a QC-related diagnostic kit.

Example 1: N-(3-(1H-imidazol-1-yl)propyl)-3',4'-dimethoxy-[1,1'-biphenyl]-2-amine (MQI-1 The synthesis of the synthesis route is as follows:

a, 3',4'-dimethoxy-[1,1'-biphenyl]-2-amine: bromoaniline (5.81 mmol, 1 equivalent), 3,4-dimethoxybenzeneboronic acid (6.98 Ment, 1.2 equivalents) and [1,1'-bis(diphenylphosphino)ferrocene]palladium(II) chloride dichloride complex (0.35 mmol, 0.06 equivalent) in a 50 ml round bottom flask Add 10 ml of dioxane and 10 ml of a 2 mol/L K ₂ CO ₃ solution, respectively, at 100 ° C for 3 h. Saturated NaCl solution was added to quench the reaction, cooled to room temperature, extracted three times with ethyl acetate, washed with saturated NaCl solution once combined, dried over anhydrous Na ₂ SO _4, the product was collected by silica gel column chromatography, in 90% yield.

b, N-(3-bromopropyl)-3',4'-dimethoxy-[1,1'-biphenyl]-2-amine: 3',4'-dimethoxy-[1 , 1'-biphenyl]-2-amine (872.32 umol, 1 equivalent), 1,3-dibromopropane (6.11 mmol, 7 equivalents) dissolved in 6 ml of anhydrous acetonitrile, anhydrous K ₂ CO ₃ (1.74 mmol) , 2 equiv) as a solid, stirred at reflux overnight, the solvent was evaporated, extracted three times with ethyl acetate and water, the organic phases were combined and washed once with saturated NaCl solution, dried over anhydrous Na ₂ SO _4, the product was collected by silica gel column chromatography to yield 48%.

c, N-(3-(1H-imidazol-1-yl)propyl)-3',4'-dimethoxy-[1,1'-biphenyl]-2-amine: imidazole (285.51 umol, 1 equivalent) dissolved in 1 ml of anhydrous acetonitrile, added to dry K ₂ CO ₃ (285.51 umol, 1 eq.), stirred at room temperature for 15 min, and added to N-(3-bromopropyl)-3', 4'- A solution of methoxy-[1,1'-biphenyl]-2-amine in 1 ml of anhydrous acetonitrile, refluxed over night, evaporated and evaporated. The aqueous Na ₂ SO _{4 was} dried, and the product was collected by silica gel column chromatography, yield 20%.

Example 2: 4'-Fluoro-N-(3-(4-methyl-1H-imidazol-1-yl)propyl)-[1,1'-biphenyl]-2-amine (MQI-31) The synthesis is based on the following route:

a, 4'-Fluoro-[1,1'-biphenyl]-2-amine: bromoaniline (5.81 mmol, 1 eq.), 4-fluorophenylboronic acid (6.98 mmol, 1.2 eq.) and [1,1' - bis(diphenylphosphino)ferrocene]palladium(II) chloride dichloride complex (0.35 mmol, 0.06 equivalent) in a 50 ml round bottom flask in 10 ml dioxane and 10 ml 2 mol /LK ₂ CO ₃ solution, stirring at 100 ° C for 3 h, adding a saturated NaCl solution to quench the reaction, extracting three times with ethyl acetate, washing once with saturated NaCl solution, drying over anhydrous Na ₂ SO ₄ , and collecting the product by silica gel column chromatography. The yield was 94%.

b, N-(3-Bromopropyl)-4'-fluoro[1,1'-biphenyl]-2-amine: 4'-fluoro-[1,1'-biphenyl]-2-amine (534.15 umol, 1 eq.) and 1,3-dibromopropane (3.74mmol, 7 eq) was dissolved in 4ml of dry acetonitrile was added anhydrous K ₂ CO ₃ (1.74mmol, ₂ eq) as a solid, stirred at reflux overnight, the solvent was evaporated Ethyl acetate and water were extracted three times. The organic phases were combined and washed once with a saturated NaCI solution, dried over anhydrous Na ₂ SO ₄ , and the product was collected by silica gel column chromatography.

c, 4'-Fluoro-N-(3-(4-methyl-1H-imidazol-1-yl)propyl)-[1,1'-biphenyl]-2-amine: 4-methylimidazole 324.48 umol, 1 equivalent) was dissolved in 1 ml of anhydrous acetonitrile, anhydrous K ₂ CO ₃ (285.51 umol, 1 eq.) was added, stirred at room temperature for 15 min; and N-(3-bromopropyl)-4'- obtained in b above was added. A solution of fluoro[1,1'-biphenyl]-2-amine in 1 ml of anhydrous acetonitrile was stirred at reflux overnight, then evaporated and evaporated, th ₂ SO _{4 was} dried, and the product was collected by silica gel column chromatography.

Example 3: QC inhibitor test for QC enzyme inhibition activity

The schematic diagram of the QC enzyme inhibition activity test is shown in Figure 1. Enzyme activity assay was performed in a 96-well microtiter plate using 200 ul of pH 8.0 Tris buffer system: 0.3 mM NADH, 2.0 mM freshly prepared Gln-Gln, 14 mM alpha-ketoglutarate, 30 U/ml glutamate dehydrogenase, 50 mM Tris, pH 8.0 buffer, finally added 0.28μM recombinant human QC protein and a mixture of inhibitors of different concentrations, after shaking for 30 seconds, dynamically change the absorption value of NADH at 340nm wavelength for 15min at 25 °C with a microplate reader. The data was collected every 30 seconds, and the IC ₅₀ value of the inhibitor against QC enzyme activity was calculated according to the test results. The test results for different QC inhibitors are shown in Table 1, wherein the structural formula of the QC inhibitor is:

Table 1

In summary, the present invention provides a glutaminyl cyclase inhibitor (QC inhibitor) which is designed according to the crystal structure of the active center of the target enzyme protein and exhibits good selectivity and specificity. It has higher drug-forming properties; its parent structure is richer, water-soluble, transmembrane performance and activity higher.

It is to be understood that the application of the present invention is not limited to the above-described examples, and those skilled in the art can make modifications and changes in accordance with the above description, all of which are within the scope of the appended claims.

Claims

A glutaminyl cyclase inhibitor characterized by the following structural formula:

Wherein, the A unit is a benzene ring, a six-membered heteroaryl ring, a five-membered heteroaryl ring, a seven-membered aromatic ring, a naphthalene, an anthracene, a naphthoquinone or a polyaromatic ring system, and R 1 is a hydrogen, a linear alkyl group, or a branched chain. One of an alkyl group, an alkoxy group, a halogen, a carboxyl group, a nitro group, a sulfonic acid group, an amine group, a phosphate group, and a substituent thereof;

The B unit is a benzene ring, a six-membered heteroaryl ring, a five-membered heteroaryl ring, a seven-membered aromatic ring, a naphthalene, an anthracene, a naphthoquinone or a polyaromatic ring system, and R 2 is a hydrogen, a linear alkyl group, or a branched alkane. One of a group, an alkoxy group, a halogen, a carboxyl group, a nitro group, a sulfonic acid group, an amine group, a phosphate group, and a substituent thereof;

In the C unit, R 3 on the imidazole ring is one of hydrogen, a linear alkyl group, a branched alkyl group, an alkoxy group, a halogen, an amine group, and a substituent thereof.
The glutaminyl cyclase inhibitor according to claim 1, wherein the B unit and the C unit are ortho, meta or para at the linking position of the A unit.
The glutaminyl cyclase inhibitor according to claim 1, wherein in the same compound, the parent core structures of the A unit and the B unit are the same or different, and the structures of R 1 and R 2 are the same or Not the same.
The glutaminyl cyclase inhibitor according to claim 1, wherein R 1 is a monosubstituted or a polysubstituted at a different position.
The glutaminyl cyclase inhibitor according to claim 1, wherein R 2 is a monosubstituted or a polysubstituted at a different position.
The glutaminyl cyclase inhibitor according to claim 1, wherein R 3 is a monosubstituted or polysubstituted at a different position.
The glutaminyl cyclase inhibitor according to claim 1, wherein the substitution position of R 3 is at the 2, 4 or 5 position of the imidazole ring.
The glutaminyl cyclase inhibitor according to claim 1, wherein in the C unit, the carbon atom in the alkyl chain is n = 1-4.